The present invention relates to the use of a treated Spirulina compound as a biological agent for the control, prevention and eradication of infections and skin defects in vertebrates and mammals. The present invention also relates to a method of reducing, or avoiding the formation of, skin defects in vertebrates, and particularly mammals.
The use of Spirulina or Spirulina extracts for care of skin and to improve the appearance of healthy skin is known. Examples of such can be found in EP 629397, FR 2609246, CN 1102069(Abstract), CN 1105566 (Abstract) amongst others. The application of Spirulina or Spirulina extracts to hair or in a shampoo is also known, for example EP 619112.
Spirulina also been found to have anti-viral abilities, as can be seen in U.S. Pat. No. 5,585,365 (Hayashi et al.) A hot water extract of Spirulina from which a calcium polysaccharide was purified was disclosed. The purified extract was effective for the treatment of viral diseases.
In addition Spirulina has been reported as having biological activity for example: to lower blood sugar levels (in diabetes); to lower blood cholesterol; and other effects (Hayashi et al). Other biological activity for Spirulina has been disclosed, for example: by Amirani (Chemical Abstracts, 91:169142n). This disclosure is to the anti-bacterial activity for Spirulina. In U.S. Pat. No 4,886,756 Kawamura discloses a restriction endonuclease from Spirulina. In J 6303391 (Abstract) a method is disclosed for producing a physiologically active substance from Spirulina.
However, in each of the above described types of biological activity of Spirulina, there is no consistency in the method of production of the Spirulina or Spirulina extract. Across the disclosures there is no reference as to whether a particular type of extract or compound produces a particular type of biological activity best.
It is an object of the present invention to overcome this absence by the use of a treated Spirulina compound. It is a further object of one aspect of the present invention to provide an improved method of biological control whereby the vigor of the agent can be controlled by the exclusion of visible light.
Another object of the present invention is the provision of a method of producing a biologically active, treated Spirulina compound which can be used effectively for any of the above described biological activity. A further object of the invention is the provision of a cost effective method as a useful choice to those presently existing.
The present invention provides a method of preparation of a treated Spirulina compound, said method comprising the steps of:
rehydrating desiccated Spirulina cells in a rehydrating solution containing
sodium bicarbonate for between 1 to 12 hours; stressing the culture by one of nutrient diminution or partial desiccation; and
freeze drying the compound.
Preferably the freeze drying produces an axenic compound.
The present invention also provides a method of biological control to inhibit and suppress the growth and effects of biological contaminants by the use of a treated Spirulina compound, said method comprising the steps of: preparing said treated Spirulina compound by the method described above, and
the application of said treated Spirulina compound on or adjacent a target site, said site being selected from the group consisting of:
an external result of the operation of the biological contaminant on a vertebrate, wherein said external result is selected from the group consisting of a papule, a stomatitis, a blister, a lesion, a pustule and a combination thereof; and old target sites where scar tissue remains.
The present invention further provides a method for the repair of skin defects in mammals; said method including the steps of: preparing said treated Spirulina compound by the method described above; and distributing said compound over a target site containing a skin defect; wherein said skin defect is selected from the group consisting of: scars, pits, reddened areas, cracks, burns, blisters, psoriasis, eczema, scaling, wrinkles and a combination thereof.
The present invention further provides a method for the avoidance of skin defects in mammals; said method including the steps of: preparing said treated Spirulina compound by the method described above; and distributing the compound over a target site which could become a skin defect; wherein said skin defect is selected from the group consisting of: scars, pits, reddened areas, cracks, burns, blisters, psoriasis, eczema, scaling, wrinkles and a combination thereof.
Preferably, the method further includes a means to control the vigor of said agent, the means being visible light, and wherein the agent is applied as a liquid culture spray.
Preferably, said compound is a liquid culture and is applied as a spray on or adjacent the target site. Alternatively, said compound is a powder of desiccated cells, evenly distributed over the target site by shake or puffer application in respect of an unicellular or multicellular organism and/or mixing in respect of a growth substrate. Optionally, said compound (liquid culture or powder form) is carried in a base such as a gel or cream for topical application to either an epidermal lesion of a human or animal, or impregnated in a surgical dressing.
By way of example only, preferred embodiments of the present invention are described in detail, with reference to a series of Examples.
In the description of the preferred embodiments, the terms xe2x80x9cfungal contaminantsxe2x80x9d, xe2x80x9ccompanion organismsxe2x80x9d and xe2x80x9cbiological agentxe2x80x9d have the meanings given below:
Fungal contaminantsxe2x80x94Deuteromycota and Ascomycota which produce conidia or asci respectively;
Companion Organismsxe2x80x94the target partner entering into a mutualistic association with the biological agent;
Biological agentxe2x80x94viable culture of cyanobacterial cells of the genera, Spirulina, possessing both photosynthetic and nitrogen fixation, metabolic activity.
Experiments were carried out using the following:
Fungal Contaminants: Hypocreaceaexe2x80x94Trichoderma viride Canterbury CTV 1; Trichophyton sp. or Epidernophyton sp.xe2x80x94causal agents of tinea pedis dermatocycosis (athlete""s foot) in humans. Treatment of the infection was conducted in situ.
The fungal contaminants are available through the Department of Zoology, University of Canterbury, New Zealand.
Companion Organismsxe2x80x94Basidiomycota: Pleurotus pulmonarius Canterbury CPP 1 (Oyster mushroom); or human in vivo.
Biological Agent: Spirulina platensis (Earthrise Farms, Mass Culture Facilities, California).
Compound Preparation:
Desiccated Spirulina platensis cells were rehydrated in 0.84 w/v sodium bicarbonate (pH 8.5) for approximately 12 hours, preferably under illumination (8-80 Watts/m2).
Optionally, the rehydration solution may further contain 0.04-0.25 w/v sodium sulphide to inhibit bacterial growth.
The culture to be stressed by nutrient diminution or partial desiccation to yield a resting stage.
Freeze drying the compound produced an axenic powder. Alternatively, commercially available desiccated S. platensis cells (Earthrise Farms) were utilized.
Secondary Infection of Human Peripheral Epidermal Tissue:
Superficial burns and acne pustules were treated in situ. Indications of microbial infection were the formation of pustules (pimple), peripheral inflammation (rubor) and swelling (tumor) at the site of trauma (acne lesion or burn).
Laboratory Trials: